Purification method of deproteinized calf blood serum extract

A calf serum and protein-removing technology, applied in the biological field, can solve the problems of low product purity, potential safety hazards, and low activity, and achieve the effects of improving purity and biological activity, ensuring safety, and broad application prospects

Inactive Publication Date: 2012-12-05
FUDAN UNIV
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Problems solved by technology

However, the products extracted by the above method have low purity and low activity, and no virus inactivation has been carried out, so the biological safety of the product

Method used

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Examples

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Embodiment 1

[0011] Embodiment 1, extract calf serum deproteinized extract

[0012] 1. Extraction and purification method

[0013] (1) Take venous blood from young cattle and separate the serum:

[0014] Under sterile conditions, collect venous blood from calves less than 6 months old, put it in a refrigerator at 4°C overnight, let it naturally separate part of the serum, and centrifuge the rest at 4000rpm, 4°C for 15 minutes under sterile conditions to separate the serum spare.

[0015] (2) Ethanol removes protein:

[0016] Add 96% ethanol to the serum according to the volume ratio of ethanol and serum of 2:1, stir constantly, let it stand for 60 minutes to denature and precipitate the protein, centrifuge (4000 rpm, 10 minutes) and leave the supernatant to discard the precipitate.

[0017] (3) In addition to ethanol:

[0018] Use a vacuum rotary evaporator to remove ethanol under reduced pressure at 37°C and 1 atmosphere.

[0019] (4) Coarse filtration:

[0020] Roughly filter with ...

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Abstract

The invention belongs to the technical field of biotechnology and provides a purification method of deproteinized calf blood serum extract. The purification method includes collecting calf venous blood, separating to obtain serum, deproteinizing with ethanol, decompressing to remove the ethanol, adding compound protease for hydrolization, centrifugally separating after hydrolization and reserving supernatant, and ultra-filtrating the supernatant; subjecting ultrafiltrate to gel chromatography with Sephadex G-15 for desalting, and collecting specific absorption elution at 280nm position; performing gel chromatographic separation with DEAE-Sephadex-50, and collecting specific absorption elution at the second 280nm position; and filtering with microporous membranes and inactivating viruses by ultraviolet irradiation so as to obtain the deproteinized calf blood serum extract. A great many of inactive ingredients are removed, molecular weight range of products is reduced, probability of adverse reactions is reduced, product purity, safety and bioactivity are improved, and the purification method is widely applicable.

Description

technical field [0001] The invention belongs to the field of biotechnology, and relates to a method for extracting and purifying calf serum deproteinized extract. Background technique [0002] Calf serum deproteinized extract (protein-free calf blood extract) is a small molecule biologically active substance obtained from 1-6 month old calf serum after deproteinization, containing small molecule peptides, amino acids, keto acids and other substances. Its main pharmacological effect is to enhance the uptake and utilization of oxygen and glucose by cells, and has the physiological effect of improving cell metabolism. It has little toxicity and is widely used in clinical practice. However, a small amount of impurities contained in the deproteinized extract of calf serum, such as inactive components such as trace proteins and even viruses, will still bring some side effects, which also narrows the scope of application. Therefore, further extraction of the deproteinized extract o...

Claims

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Application Information

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IPC IPC(8): A61K35/16A61P43/00
Inventor 陈放李如伟
Owner FUDAN UNIV
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